Anti-MCT8 抗体 [EPR26257-174] (BSA and Azide free) (ab302707)

All lanes : Anti-MCT8 antibody [EPR26257-174] (ab302706) at 1/1000 dilution

Lane 1 : Mouse kidney tissue lysate
Lane 2 : Mouse liver tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Rat liver tissue lysate
Lane 5 : Rat spleen tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Performed under non-reducing conditions.

Predicted band size: 59 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer and concentration: 5% NFDM/TBST. 

Samples are non-boiled as boiling may cause protein aggregates.
Negative control: spleen (PMID:9545634). 

Exposure time: Lane 1: 48 seconds, Lane 2-5: 81 seconds

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling MCT 8 with ab302706 at 1/2000 dilution (0.252 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining in human liver was observed (PMID:19641107, PMID:12871948 ). The section was incubated with ab302706 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling MCT 8 with ab302706 at 1/5000 dilution (0.101 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining in mouse kidney was observed (PMID: 19641107, PMID: 19996182). The section was incubated with ab302706 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling MCT 8 with ab302706 at 1/5000 dilution (0.101 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining in rat liver was observed ( (PMID:19641107, PMID:12871948). The section was incubated with ab302706 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins as used. 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells transfected with a MCT8 expression vector containing a his tag tissue labeling MCT 8 with ab302706 at 1/5000 dilution (0.101 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in (A) HEK-293T cells transfected with a MCT8 expression vector containing a his tag was observed and no staining in (B) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab302706 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling MCT 8 with ab302706 at 1/2000 dilution (0.252 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining in human spleen. The section was incubated with ab302706 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.  

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MCT 8 with ab302706 at 1/5000 dilution (0.101 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining in mouse spleen (PMID: 9545634). The section was incubated with ab302706 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labeling MCT 8 with ab302706 at 1/100 dilution (5.03 µg/mL) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 diluion (2 µg/mL) (Green). Positive staining on mouse liver is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Primary dilutent was used instead of primary antibody, follwed by secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 diluton (2 µg/mL). 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen (fresh) tissue labeling MCT 8 with ab302706 at 1/100 dilution (5.03 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control: no staining on mouse spleen (PMID: 25409824) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Primary dilutent was used instead of primary antibody, follwed by secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 diluton (2 µg/mL). 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling MCT 8 with ab302706 at 1/100 dilution (5.03 µg/mL) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/mL) dilution (Green). Positive staining on rat liver is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Primary diluent was used instead of primary antibody, followed by secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL). 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat spleen (fresh) tissue labeling MCT 8 with ab302706 at 1/100 (5.03 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Negative control: no staining on rat spleen (PMID: 25409824) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.<\p>

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This data was developed using 302706, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labeling with at 1/ 500 dilution (1.006 μg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing positive staining in mouse primary neuron. Anti-MAP2 mouse monoclonal antibody ab11267 (Alexa Fluor^® 594) was used to counterstain microtubuls at 1/500 dilution (4 μg/mL), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (2 μg/mL)(Red). The Nuclear counterstain was (Blue).

The negative controls are as follows: 

-ve control 1: ab302706 at 1/500 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1:1000 dilution (2 μg/mL)

-ve control 2: ab11267 (Anti-MAP2 mouse monoclonal antibody) at 1:500 dilution (4 μg/mL)  followed by ab150081 at 1:1000 dilution (2 μg/mL)

Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. 

This data was developed using 302706, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labeling with at 1/ 500 dilution (1.006 μg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing positive staining in rat primary neuron. Anti-MAP2 mouse monoclonal antibody ab11267 (Alexa Fluor^® 594) was used to counterstain microtubuls at 1/500 dilution (4 μg/mL), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (2 μg/mL)(Red). The Nuclear counterstain was (Blue).

The negative controls are as follows: 

-ve control 1: ab302706 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1:1000 dilution (2 μg/mL)

-ve control 2: ab11267 (Anti-MAP2 mouse monoclonal antibody) at 1:500 dilution (4 μg/mL)  followed by ab150081 at 1:1000 dilution (2 μg/mL)

Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized mouse primary neural/glia cells labeling MCT 8 with ab302706 at 1/50 dilution (1µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

Intracellular Ffow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized rat primary neural/glia cells labeling MCT 8 with ab302706 at 1/50 dilution (1 µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

MCT 8 was immunoprecipitated from 0.35 mg mouse liver tissue lysate with ab302706 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302706 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse liver tissue lysate 10 µg (Inset)

Lane 2: ab302706 IP in mouse liver tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302706 in mouse liver tissue lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 36 seconds

Observed MW (kDa): 60

 

This data was developed using ab302706, the same antibody clone in a different buffer formulation.

MCT 8 was immunoprecipitated from 0.35 mg Rat liver tissue lysate with ab302706 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302706 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Rat liver tissue lysate 10 µg (Inset)

Lane 2: ab302706 IP in rat liver tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302706 in rat liver tissue lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 36 seconds

Observed MW (kDa): 60